State Key Laboratory of Stem Cell and Reproductive Biology, Institute of Zoology, Chinese Academy of Sciences, Beijing, 100101, China; University of Chinese Academy of Sciences, Beijing, 100049, China.
State Key Laboratory of Stem Cell and Reproductive Biology, Institute of Zoology, Chinese Academy of Sciences, Beijing, 100101, China.
Biochem Biophys Res Commun. 2020 Jun 18;527(1):8-14. doi: 10.1016/j.bbrc.2020.04.078. Epub 2020 Apr 24.
Oocyte meiotic maturation failure and unfaithful chromosome segregation are major causes for female infertility. Here, we showed that CENP-W, a relatively novel member of the kinetochore protein family, was expressed in mouse oocytes from the germinal vesicle (GV) to metaphase II (MII) stages. Confocal microscopy revealed that CENP-W was localized in the germinal vesicle in the GV stage, and then became concentrated on kinetochores during oocyte maturation. Knockdown of CENP-W by specific siRNA injection in vitro caused kinetochore-microtubule detachment, resulting in severely defective spindles and misaligned chromosomes, leading to metaphase I arrest and failure of first polar body (PB1) extrusion. Correspondingly, spindle assembly checkpoint (SAC) activation was observed in CENP-W knockdown oocytes even after 10h of culture. Our results suggest that CENP-W acts as a kinetochore protein, which takes part in kinetochore-microtubule attachment, thus mediating the progression of oocyte meiotic maturation.
卵母细胞减数分裂成熟失败和染色体分离错误是女性不孕的主要原因。在这里,我们表明,着丝粒蛋白家族的一个相对新颖的成员 CENP-W,在从生发泡(GV)到中期 II(MII)阶段的小鼠卵母细胞中表达。共聚焦显微镜显示 CENP-W 在 GV 期位于生发泡中,然后在卵母细胞成熟过程中集中在动粒上。通过特异性 siRNA 注射在体外敲低 CENP-W 会导致动粒-微管分离,从而导致严重缺陷的纺锤体和染色体排列不齐,导致中期 I 阻滞和第一极体(PB1)排出失败。相应地,即使在培养 10 小时后,CENP-W 敲低卵母细胞中也观察到纺锤体组装检查点(SAC)的激活。我们的结果表明,CENP-W 作为一种着丝粒蛋白,参与着丝粒-微管的附着,从而介导卵母细胞减数分裂成熟的进行。